东方蜜蜂DNA随机扩增多态及其遗传分化研究

本研究对云南及马来西亚的37个东方蜜蜂样本进行随机增多态DNA分析，从20个引物中筛选出11个引物，其中9个引物扩增出多态带。共检测到66条扩增片段，其中56条为多态带。用UPGMA聚类方法构建的分子系统树显示，云南的样本、马来西亚的样本各自分别聚在一起，说明两个样本间遗传差异较大，群体之间存在着遗传分化。但就云南的32个个体而言，虽然聚类图中大多数采用自同一地区的样本聚在一起，但也存在一定交叉，提示云南地理群体间近期可能存在一定的基因流。     

4种体色柞蚕品种遗传关系的RAPD分析

利用随机扩增多态DNA(RAPD)标记技术,对分别属于4种体色系统的13个柞蚕品种进行了基因组DNA多态性分析。利用筛选出的33个随机引物扩增出265条DNA带,其中多态性带227条,占85.66%。根据扩增结果计算品种间的单匹配相似系数,用Ne ighbor-Join ing法构建了聚类树状图。4种体色13个品种并没有按所属体色系统聚在一起,即品种的遗传聚类划分与体色之间的相关性不大,表明柞蚕体色并不能真实反映品种间的遗传关系。     

AFLP标记检测大足黑山羊地方类群遗传多样性的研究

利用AFLP标记技术,从15对引物中选取11对扩增效果较好的引物,对17只大足黑山羊个体DNA进行扩增。结果共扩增出766个条带,其中59条为多态条带,平均每对引物扩增出69．6个条带,平均多态率为5．36％.     

野生垂穗披碱草种质遗传多样性的SRAP研究

用SRAP标记对采集自中国四川、西藏、甘肃、青海、新疆的68份垂穗披碱草种质进行了多态性分析,获得下述结果,1)用20对引物共扩增出495条带,平均每对引物扩增出24.75条,其中多态带425条,平均每对引物21.25条,多态率达85.39%。材料间遗传相似系数变化范围为0.374~0.997,平均值为0.745。这些结果说明,供试垂穗披碱草材料具有较为丰富的遗传多样性。2)对所有材料的聚类分析和主向量分析发现,在GS值为0.77的水平上供试材料可聚成5个类群,大部分来自相同或相似生态地理环境的材料聚为一类,表明供试材料的聚类和其生态地理环境间有一定的相关性。     

玳瑁遗传多样性的RAPD分析

应用RAPD技术分析了玳瑁的遗传多样性。用20个随机引物对中国南海海域玳瑁7个个体的基因组DNA进行了PCR扩增,共扩增出1 351条DNA片段,平均每个个体扩增出193条条带。在检测到的193条条带中,多态性条带为69条,多态性条带百分比为35.8%,条带大小在200 bp～3 000 bp之间,7个个体间遗传距离为0.082 9～0.1813,平均遗传距离为0.132 7±0.029 9,表明中国南海海域玳瑁的遗传多样性水平较低,应加强该区域玳瑁种质资源的保护。采用类平均聚类法（NJTREE）构建了7个个体相互关系的分子聚类图,表明该7个玳瑁个体没有形成种群的分化。     

贵州黑山羊遗传结构的RAPD分析

用27条多态性引物对15份贵州黑山羊个体基因组进行RAPD分析结果共扩增出258条带,其中多态带为119条,多态频率在0-77．78％之间,平均多态频率为46．12％．单个引物获得的扩增带数在4-14条之间,平均每条引物扩增出9．56条带、扩增带分子量范围为230bp-2800bp、用Nei氏公式计算品种内个体间的遗传相似性指数平均为0．9023结果表明：贵州黑山羊个体间的遗传变异较大,具有较丰富的遗传结构。     

贵州白山羊遗传结构的RAPD分析

用27条多态性引物对15份贵州白山羊个体基因组进行RAPD分析。结果共扩增出268条带，其中多态带为112条，多态频率在0～72．73％之间，平均多态频率为41．79％。单个引物获得的扩增带数在4～16条之间，平均每条引物扩增出9．92条带，扩增带分子量范围在230～2800bp。Nei氏公式计算品种内个体阃遗传相似指数平均为0．9156。结果表明：贵州白山羊个体间遗传变异较小，具有较高的遗传稳定性。     

野生马蹄金种质遗传多样性的SRAP研究

为评价野生马蹄金种质的遗传多样性,从80对SRAP引物中筛选出19对多态性高、稳定性好的引物,用于研究21份马蹄金材料,获得如下结果:1)19对引物共扩增出212条带,平均每对引物扩增出11.1条,其中多态性条带156条,平均每对引物8.2条,多态率为73.58%。材料间遗传相似系数变化范围为0.444~0.980,平均值为0.711 2。结果表明,供试马蹄金材料具有较为丰富的遗传多样性。2)对所有材料的聚类分析发现,在GS值为0.78处所有供试材料可聚为3个类群,大部分来源地相同或生态环境相似的材料聚为一类,表明供试材料的聚类和生态地理环境具有一定的相关性。     

野生马蹄金种质遗传多样性的SRAP研究

为评价野生马蹄金种质的遗传多样性,从80对SRAP引物中筛选出19对多态性高、稳定性好的引物,用于研究21份马蹄金材料,获得如下结果:1)19对引物共扩增出212条带,平均每对引物扩增出11.1条,其中多态性条带156条,平均每对引物8.2条,多态率为73.58%。材料间遗传相似系数变化范围为0.444~0.980,平均值为0.7112。结果表明,供试马蹄金材料具有较为丰富的遗传多样性。2)对所有材料的聚类分析发现,在GS值为0.78处所有供试材料可聚为3个类群,大部分来源地相同或生态环境相似的材料聚为一类,表明供试材料的聚类和生态地理环境具有一定的相关性。     

冰草属植物种质资源遗传多样性的ISSR分析

用ISSR标记对来自国内外的33份冰草材料的遗传多样性进行了检测。从93条ISSR引物中共筛出11条能扩增出清晰条带并具有多态性的引物,33个样品DNA共获得84个扩增位点,其中多态性位点59个,平均每个引物扩增位点为7.64个。品种间遗传相似系数在0.083～0.706,表现出丰富的遗传多样性。利用UPGMA聚类分析,以遗传相似系数0.52为界限,33份材料划分为4类,聚类基本符合地理来源相近的材料聚为一类,呈现出一定的地域性分布规律。     

Phase I evaluation of CCNU (lomustine) in tumor-bearing cats

1-(2-Chloroethyl)3-cyclohexyl-1-nitrosourea (CCNU) is an alkylating agent in the nitrosourea subclass. A prospective evaluation of CCNU was done to determine the maximally tolerated dosage of CCNU in tumor-bearing cats. Response data were obtained when available. Twenty-five cats were treated with CCNU at a dosage of 50-60 mg/m3 body surface area. Complete hematologic data were available for 13 cats. Neutropenia was the acute dose-limiting toxicity. The median neutrophil count at the nadir was 1,000 cells/microL (mean, 2,433 cells/microL; range, 0-9,694 cells/microL). The time of neutrophil nadir was variable, occurring 7-28 days after treatment, and counts sometimes did not return to normal for up to 14 days after the nadir. Based on these findings, a 6-week dosing interval and weekly hematologic monitoring after the 1st treatment with CCNU are recommended. The nadir of the platelet count may occur 14-21 days after treatment. The median platelet count at the nadir was 43,500 cells/microL. No gastrointestinal, renal, or hepatic toxicities were observed after a single CCNU treatment, and additional studies to evaluate the potential for cumulative toxicity should be performed. Five cats with lymphoma and 1 cat with mast cell tumor had measurable responses to CCNU. Phase II studies to evaluate antitumor activity should be completed with a dosing regimen of 50-60 mg/m3 every 6 weeks.     

Furosemide continuous rate infusion in the horse: evaluation of enhanced efficacy and reduced side effects

Continuous rate infusion (CRI) of furosemide in humans is considered superior to intermittent administration (IA). This study examined whether furosemide CRI, compared with IA, would increase diuretic efficacy with decreased fluid and electrolyte fluctuations and activation of the renin-angiotensin-aldosterone system (RAAS) in the horse. Five mares were used in a crossover-design study. During a 24-hour period, each horse received a total of 3 mg/kg furosemide by either CRI (0.12 mg/kg/h preceded by a loading dose of 0.12 mg/kg IV) or IA (1 mg/kg IV q8h). There was not a statistically significant difference in urine volume over 24 hours between methods; however, urine volume was significantly greater after CRI compared with IA during the first 8 hours ([median 25th percentile, 75th percentile]: 9.6 L [8.9, 14.4] for CRI versus 5.9 L [5.3, 6.0] for IA). CRI produced a more uniform urine flow, decreased fluctuations in plasma volume, and suppressed renal concentrating ability throughout the infusion period. Potassium, Ca, and Cl excretion was greater during CRI than IA (1,133 mmol [1.110, 1,229] versus 764 mmol [709, 904], 102.7 mmol [96.0, 117.2] versus 73.3 mmol [65.0, 73.5], and 1,776 mmol [1,657, 2.378] versus 1,596 mmol [1,457, 1,767], respectively). Elimination half-lives of furosemide were 1.35 and 0.47 hours for CRI and IA, respectively. The area under the excretion rate curve was 1,285.7 and 184.2 mL x mg/mL for CRI and IA, respectively. Furosemide CRI (0.12 mg/kg/h) for 8 hours, preceded by a loading dose (0.12 mg/kg), is recommended when profound diuresis is needed acutely in horses.     

The Epidemiology of Gastrointestinal Nematodes of Dairy Cattle in Central Kenya

The epidemiology of H. placei and of other gastrointestinal nematodes in yearling dairy cattle was examined on two farms in Kiambu District, central Kenya during each of 13 one-month periods from April 1993 to April 1994. On each farm, 32 newly weaned dairy calves were given a single dose of albendazole and then placed on experimental pastures. Twelve of the animals were designated for bi-monthly slaughter (n = 2) and analysis of worm population characteristics and 20 were designated for blood and faecal collection and for weighing. Two parasite-free tracer calves were grazed alongside the weaner calves each month throughout the study period and were also slaughtered for analysis of worm populations. Faecal egg counts, haematological and serum pepsinogen determinations, herbage larval counts, and animal live weight changes were recorded monthly. The study revealed that Haemonchus placei, Trichostrongylus axei, Cooperia spp. and Oesophagostomum radiatum were responsible for parasitic gastroenteritis and that H. placei was the predominant nematode present in the young cattle on both farms. Faecal egg counts from resident cattle and necropsy worm counts revealed that pasture larval levels were directly related to the amount of rainfall. The total worm burdens in the animals were highest during the rainy season (March–June and October–December) and lowest during the dry seasons (July–September and January–February). The very low recovery of immature larvae of H. placei from the tracer calves indicated that arrested development is not a feature of the life cycle of this parasite in central Kenya. The maintenance of the parasite population depended on continuous cycling of infection between the host and the pasture. The agroclimatic conditions of the study area were such that, in general, favourable weather conditions for the development and survival of the free-living stages of gastrointestinal nematodes existed all year round.     

Diversity of Ectoparasites in Sheep Flocks in Sa~o Paulo,Brazil

The occurrence of ectoparasites in sheep flocks is frequently reported but seldom quantified. Sheep production used to be a predominantly family activity in the state of Sa~o Paulo (Brazil), but it began to become a commercial activity in the past decade. Thus, information about the ectoparasites existing in sheep flocks has become necessary. The present data were obtained by means of questionnaires sent to all sheep breeders belonging to the `Associaça~o Paulista de Criadores de Ovinos' (ASPACO; Sa~o Paulo State Association of Sheep Breeders). Response reliability was tested by means of random visits paid to 10.6% of the respondents. Most of the properties (89.5%) reported the presence of one or more ectoparasites. Screw-worm (Cochliomyia hominivorax) was the most frequent ectoparasite (72.5%), followed by bot fly larvae (Dermatobia hominis, 45.0%), ticks (Amblyomma cajennense) and Boophilus microplus, 31.3%) and finally lice (Damalinia ovis, 13.8%). Combined infestations also occurred, the most common one being screw-worm with bot fly larvae (36.0%) followed by bot fly larvae with ticks (13.9%), screw-worm with ticks (9.3%), bot fly larvae with lice (6.9%), and ticks with lice (5.0%). The most common triple combination was screw-worm, bot fly larvae and ticks (12.8%). Breeds raised for meat or wool were attacked by bot fly larvae and ticks more often than other breeds. Lice were only absent from animals of indigenous breeds. The relationships among these ectoparasites are discussed in terms of sheep breeds, flock size, seasonality and the ectoparasitic combinations on the host.     

Evaluation of biohydrogenation rate of canola vs. soya bean seeds as unsaturated fatty acids sources for ruminants in situ

An experiment was conducted to study disappearance of C14 to C18 fatty acids, lag times and biohydrogenation (BH) rates of C18 fatty acids of ground soya bean and canola seeds in situ. Three ruminally fistulated Dallagh sheep were used to determine ruminal BH of unsaturated fatty acids (UFAs). Differences in the disappearance of fatty acids through the bags and lag times were observed between the oilseeds. We saw that the longer the incubation time of the oilseeds in the rumen, the lower the content of C18:2 and C18:3. Significantly higher lag times for both C18:2 and C18:3 were observed in ground canola compared to ground soya bean. BH rates of C18:2 and C18:3 fatty acids in soya bean were three times higher than those of canola. These results suggest that the fatty acid profile of fat source can affect the BH of UFAs by rumen micro‐organisms. So that UFAs of canola had higher ability to escape from ruminal BH. It seems that fatty acid profile of ruminant products is more affected by canola seed compared to soya bean seed.     

Serum biochemical values and mineral element contents of tissues in yaks

Serum biochemical values and the wet weight to dry weight ratios of tissues were determined in yaks in Shandan county of Gansu province. The liver, kidney, heart and muscle contents of seven elements in yaks were also analysed. Most serum biochemical values were similar to those of cattle, camels and sheep, but the calcium concentration was considerably above the normal range for other ruminants. The liver contained the highest concentrations of copper, manganese and cobalt and the kidney of selenium, iron and calcium.Abbreviations Alb albumin - ALT alanine aminotransferase - AST aspartate aminotransferase - BUN blood urea nitrogen - Chol cholesterol - Glob globulin - -GT -glutamyl transpeptidase - IP inorganic phosphorus - TP total protein     

Evaluation of the Microbiological Status of Milk and Various Structures in Mammary Glands from Naturally Infected Dairy Cows

A knowledge of the microbiological status of milk and of the different structures in the mammary glands has great importance in elucidating the pathogenesis of mammary gland infections. The objective of this study was to evaluate the microbiological status of various structures in the mammary glands from naturally infected dairy cows following slaughter. A total of 94 samples of milk, 184 samples of mammary parenchyma, 168 samples of gland cisterns, and 168 samples of teat cisterns were collected for microbiological examination. Microorganisms were detected in 59.9% of all samples, 67.0% of the milk samples, 70.1% of the mammary parenchymas, 55.9% of the gland cisterns and 48.8% of the teat cistern samples. When all samples were considered, coagulase-negative Staphylococcus were the most prevalent (35.7%) followed by coagulase-positive Staphylococcus (12.2%), Corynebacterium bovis (2.4%), Prototheca sp. (1.9%), and Streptococcus dysgalactiae (1.5%). There was a significantly higher occurrence of microorganisms in the milk and mammary parenchyma compared to the gland cisterns and teat cisterns. This revised version was published online in July 2006 with corrections to the Cover Date.     

Development and survival of free-living stages of gastrointestinal nematodes of sheep and goats on pasture in the Nigerian derived savanna

Four grass plots were sequentially contaminated with goat faeces containing known numbers of unembryonated eggs of predominantly Haemonchus contortus and Trichostrongylus spp. between October 1982 and April 1983. Four other plots were similarly contaminated with sheep faeces between February and May 1987. An additional plot was repeatedly contaminated with sheep faeces from February to April 1987. Populations of free-living stages in faeces and of infective larvae (L₃) in the herbage were subsequently monitored until the end of April and June of 1983 and 1987 respectively. During February and May 1987 two control cultures of sheep faeces were incubated in the laboratory at 25°C–30°C and at a constant temperature of 50°C and the free-living development was also monitored. L₃ developed very readily in the faeces cultured at 25°C–30°C and in those spread on a grass plot in October, at the end of the wet season, but developed less on the plot contaminated in May at the start of the wet season. Worm eggs in faeces deposited on plots during the hot dry season (December to April) or incubated at 50°C died and disintegrated after 24–48 h exposure to the high environmental temperatures. The results indicate that it is unlikely that gastrointestinal nematodes of sheep and goats can develop or survive on open pasture during the dry season in the Nigerian derived savanna zone.